Fig. 5: SPOP promotes β-catenin degradation and ubiquitination. | Cell Death Discovery

Fig. 5: SPOP promotes β-catenin degradation and ubiquitination.

From: O-GlcNAcylation of SPOP regulates colorectal cancer progression and ferroptosis by mediating β-catenin degradation

Fig. 5

A The expression of β-catenin in SPOP-overexpressing HCT116 and DLD1 cells was analyzed by WB after treatment with MG132 (10 μM, 6 h). B HCT116 cells transfected with shSPOP were treated with CHX, and samples were harvested at multiple time points for WB. C Total ubiquitination levels of β-catenin in SPOP-knockdown HCT116 and DLD1 cells transfected with the corresponding plasmids (48 h) and treated with MG132 (10 μM, 6 h). D 293 T cells were transfected with MYC-β-catenin, HA-Ub and Flag-SPOP. The cell lysates were subjected to IP analysis. E Sequence comparison of β-catenin with the SBC motif in known SPOP substrates. F 293 T cells were transfected with MYC-β-catenin-SBC1, MYC-β-catenin-SBC2 and Flag-SPOP. The cell lysates were analyzed via IP. G Ubiquitination assays via IP in 293 T cells transfected with MYC-tagged β-catenin SBC mutants, HA-Ub, and Flag-SPOP. H Analysis of β-catenin ubiquitination in 293 T cells co-expressing MYC-β-catenin, various HA-Ub linkage mutants (K27, K48, K63), and Flag-SPOP using IP. I IP analysis of ubiquitination in 293 T cells transfected with K-to-R β-catenin mutants (K170R, K233R, K508R), HA-Ub, and Flag-SPOP.

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